Hubbell, Jeffrey AlanSpatz, JoachimVishwakarma, Medhavi2017-03-092017-03-092017-03-09201710.5075/epfl-thesis-7392https://infoscience.epfl.ch/handle/20.500.14299/135101urn:nbn:ch:bel-epfl-thesis7392-9Collective migration is a central event involving coordinated movement of several individuals. One of the critical events during collective migration is the emergence of leading individuals, who provide directional guidance for the group movement. While on the move, animals select their leaders by a collective decision making process, which require active participation of the followers. On the contrary, the prevalent view on collective cell migration, especially in the context of epithelial cells during wound healing, assumes a hierarchical leader-follower organization and belittles the contribution of follower cells in choosing or regulating the leaders. Furthermore, how the dynamics of cells located at the wound margin evolve as the wound heals, remains illusive. Here, we report and analyse distinct phases of collective migration during wound closure and demonstrate how cellular-level shared decision-making process and collective mechanical dynamics influence selection, regulation and kinematics of leader cells in these phases. We found that in the preparatory phase, before the initiation of migration (Phase 0), the selection of leader cells at the epithelial wound margin depends on the pre-migratory dynamics of the follower cells situated immediately behind the future leaders. Long before the prospective leaders actually start displaying their phenotypic peculiarities, cells behind them manifest stochastic augmentations in the traction forces and monolayer stresses, and display large perimeter-to-area ratio indicating a local unjamming in the followers much before the leaders are selected. Further, introducing an unjammed or fluidic follower at the back stimulates leader cell formation at the margin thereby indicating the role of collective bulk dynamics in leader cell selection. Interestingly, the length upto which cells cooperatively join forces, corresponds very well with the distance between the two emerging leaders and this mechano-biological control remains preserved even in the presence of geometric bias or physiological levels of chemical cue at the interface. Immediately after the initiation of migration (Phase 1), leaders show their distinct phenotypes and drive the cellular outgrowths. In this phase, pluricellular actin belt at the margin regulates the fraction of marginal leaders, which therefore remains unchanged, while the number of followers per leader increases with time. As the migration progresses, fraction of leader cells increases while the latter settles to a steady level set again by the length scale of cell-cell force transmission (Phase 2). Any perturbations in mechanical forces that modifies the force correlation lengths, invariably enforces a change in the number of followers per leader thereby modifying the time required to transit from one phase to the other. Furthermore, orientation of focal adhesions and persistence of cellular motions also display this phase specific behaviour. Together, these findings provide a novel system insight into collective cell migration and indicate integrative leader-follower interactions during wound closure. Given the physiological and pathological importance of leader cell formation in epithelial wound healing, in organogenesis and in metastatic migration of cancer cells, the system-view that the results offer here is anticipated to have to a long-standing impact on the design and discovery of avant-garde therapeutic strategies in future.encollective cell migrationepithelial cellswound healingwound marginleader cellsnon-leader cellsfollower cellsdynamic heterogeneityforce-correlation lengthcollective dynamicsjammed cellsunjammed cellspropensity of leaderspersistence of migrationmechano-transductionfocal-adhesionsmicro-patterningtraction-force microscopymonolayer-stress microscopyreflection interference contrast microscopythesis::doctoral thesis